LOCUS Exported 4215 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Synthetic promoter responsive to gRNA1. ACCESSION . VERSION . KEYWORDS P1-EYFP-pA (Construct 5) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4215) AUTHORS Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK TITLE Multiplexed and Programmable Regulation of Gene Networks with an Integrated RNA and CRISPR/Cas Toolkit in Human Cells. JOURNAL Mol Cell. 2014 May 14. pii: S1097-2765(14)00355-4. doi: 10.1016/j.molcel.2014.04.022. PUBMED 24837679 REFERENCE 2 (bases 1 to 4215) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4215 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(468..1328) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1091..1110 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1329..1433) /gene="bla" /label=AmpR promoter rep_origin 1460..1915 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1547..1566) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1757..1778 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 2046..2094 /note="synthetic polyadenylation signal" misc_feature 2108..2199 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 2148..2167 /label=RVprimer3 /note="pGL3 vector, forward primer" CDS 2649..3353 /codon_start=1 /product="Phialidium yellow fluorescent proteim" /label=PhiYFP /note="mammalian codon-optimized" /translation="MSSGALLFHGKIPYVVEMEGDVDGHTFSIRGKGYGDASVGKVDAQ FICTTGDVPVPWSTLVTTLTYGAQCFAKYGPELKDFYKSCMPDGYVQERTITFEGDGNF KTRAEVTFENGSVYNRVKLNGQGFKKDGHVLGKNLEFNFTPHCLYIWGDQANHGLKSAF KICHEIAGSKGDFIVADHTQMNTPIGGGPVHVPEYHHMSYHVKLSKDVTDHRDNMSLTE TVRAVDCRKTYL" polyA_signal 3379..3500 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3390..3409) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3444..3463 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3753..3770) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3924..297) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4004..4023) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 61 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 121 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 181 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 241 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 301 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 361 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 421 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 481 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 541 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 601 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 661 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 721 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 781 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 841 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 901 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 961 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 1021 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 1081 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 1141 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 1201 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 1261 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 1321 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 1381 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 1441 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 1501 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 1561 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 1621 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 1681 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 1741 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 1801 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 1861 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcccat 1921 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 1981 cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg 2041 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga 2101 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg 2161 ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgatagcta gctgaaatat 2221 taatgtcgcg actattaata ttttcaggct agccatgctt cgctacacgc gactattaat 2281 attttcaggc tagccatgct tcgctacacg cgactattaa tattttcaag ctagccatgc 2341 ttcgctacac gcgactatta atattttcag gctagccatg cttcgctaac gcgactatta 2401 atattttcag gctagtcatg cttcgctaca cgcgactatt aatattttca ggctagccat 2461 gcttcgctac acgcgactat taatattttc aggctagcca tgcttcgcta cacgcgacta 2521 ttaatatttt caggctagcc atgcttcgct acacgcgact attaatattt tcaggctagc 2581 ggggggctat aaaagggggt gggggcgttc gtcctgctat ctagcgtcgc gttgaccatg 2641 gcgccaccat gagcagcggc gccctgctgt tccacggcaa gatcccctac gtggtggaga 2701 tggagggcga tgtggatggc cacaccttca gcatccgcgg taagggctac ggcgatgcca 2761 gcgtgggcaa ggtggatgcc cagttcatct gcaccaccgg cgatgtgccc gtgccctgga 2821 gcaccctggt gaccaccctg acctacggcg cccagtgctt cgccaagtac ggccccgagc 2881 tgaaggattt ctacaagagc tgcatgcccg atggctacgt gcaggagcgc accatcacct 2941 tcgagggcga tggcaatttc aagacccgcg ccgaggtgac cttcgagaat ggcagcgtgt 3001 acaatcgcgt gaagctgaat ggccagggct tcaagaagga tggccacgtg ctgggcaaga 3061 atctggagtt caatttcacc ccccactgcc tgtacatctg gggcgatcag gccaatcacg 3121 gcctgaagag cgccttcaag atctgccacg agatcgccgg cagcaagggc gatttcatcg 3181 tggccgatca cacccagatg aataccccca tcggcggcgg ccccgtgcac gtgcccgagt 3241 accaccacat gagctaccac gtgaagctga gcaaggatgt gaccgatcac cgcgataata 3301 tgagcctgac ggagaccgtg cgcgccgtgg attgccgcaa gacctacctg taaggccggc 3361 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc 3421 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 3481 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 3541 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata 3601 aggatcgagt atgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt 3661 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg 3721 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 3781 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 3841 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 3901 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 3961 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 4021 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 4081 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 4141 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 4201 gaccgctgcg cctta //